*Protein ID (NP or XP #) or Wolbachia#: LmjF14.1370
*Organism (including strain): Leishmania major
Etiologic Risk Group (see link below):
*Background/Disease Information (sort of like the Intro to your Mini Research Write up): Leishmania major is a protozoa that causes cutaneous leishmaniasis, which harms the macrophages and dendritic immune cells. It is transmitted by the female sandfly and can cause uncomfortable skin lesions.
Essentiality of this protein:
Gene/Ortholog: Tb927.7.3620 (OG4_11028); Phenotype: significant gain of fitness in procyclic forms; Source study: alsford
Complex of proteins?: yes
Druggable Target (list number or cite evidence from a paper/database showing druggable in another organism): Homo sapiens (1)
Synthetic analogues of SB-219383. Novel C-glycosyl peptides as inhibitors of tyrosyl tRNA synthetase. (2001).
Enzyme Assay information (spectrophotometric, coupled assay ?, reagents): The effect of substrate binding on trypanosomal tyrosyl-tRNA synthetase was assayed by adding substrate or other potential ligands to 0.5 mg/ml protein in standard buffer and monitoring fluorescence over a temperature range of 20–90° C. http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3095712/
Image of protein (PyMol with features delineated and shown separately): *Amino Acid Sequence (paste as text only - not as screenshot or as 'code'):
MNTDERYKLLRSVGEECIQESELRNLIEKKPLIRCYDGFEPSGRMHIAQGIFKAVNVNKC
*NCBI Gene # or RefSeq#: 27213
*Protein ID (NP or XP #) or Wolbachia#: LmjF14.1370
*Organism (including strain): Leishmania major
Etiologic Risk Group (see link below):
*Background/Disease Information (sort of like the Intro to your Mini Research Write up): Leishmania major is a protozoa that causes cutaneous leishmaniasis, which harms the macrophages and dendritic immune cells. It is transmitted by the female sandfly and can cause uncomfortable skin lesions.
Link to TDR Targets page (if present): http://tdrtargets.org/targets/view?gene_id=27213
Link to Gene Database page (NCBI, EuPath databases -e.g. TryTryp, PlasmoDB, etc - or PATRIC, etc.): http://tritrypdb.org/tritrypdb/showRecord.do?name=GeneRecordClasses.GeneRecordClass&primary_key=LmjF14.1370
Essentiality of this protein:
Gene/Ortholog: Tb927.7.3620 (OG4_11028); Phenotype: significant gain of fitness in procyclic forms; Source study: alsford
Complex of proteins?: yes
Druggable Target (list number or cite evidence from a paper/database showing druggable in another organism): Homo sapiens (1)
Synthetic analogues of SB-219383. Novel C-glycosyl peptides as inhibitors of tyrosyl tRNA synthetase. (2001).
Bioorg Med Chem Lett 11: 711-4.
*EC#: 6.1.1.1
Link to BRENDA EC# page: http://brenda-enzymes.org/php/result_flat.php4?ecno=6.1.1.1
Enzyme Assay information (spectrophotometric, coupled assay ?, reagents): The effect of substrate binding on trypanosomal tyrosyl-tRNA synthetase was assayed by adding substrate or other potential ligands to 0.5 mg/ml protein in standard buffer and monitoring fluorescence over a temperature range of 20–90° C.
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3095712/
Sigma Aldrich Supplier
ATP disodium salt hydrate-----$ 37.20 (1mg)-----FLAAS
L-tyrosine-----$65.50 (50g)----- T3754
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC390865/pdf/pnas00363-0045.pdf
Structure Available (PDB or Homology model)
-- PDB # or closest PDB entry if using homology model: 3P0J
Current Inhibitors:
Expression Information (has it been expressed in bacterial cells): Yes, expressed in E. coli.
Purification Method: Ni-NTA affinity chromatography
Image of protein (PyMol with features delineated and shown separately):
*Amino Acid Sequence (paste as text only - not as screenshot or as 'code'):
MNTDERYKLLRSVGEECIQESELRNLIEKKPLIRCYDGFEPSGRMHIAQGIFKAVNVNKC
TAAGCEFVFWVADWFALMNDKVGGELEKIRIVGRYLIEVWKAAGMDMDKVLFLWSSEEIT
SHADTYWRMVLDIGRQNTIARIKKCCTIMGKTEGTLTAAQVLYPLMQCCDIFFLKADICQ
LGLDQRKVNMLAREYCDLIGRKLKPVILSHHMLAGLKQGQAKMSKSDPDSAIFMEDTEED
VARKIRQAYCPRVKQSASAITDDGAPVATDDRNPVLDYFQCVVYARPGAAATIDGTTYAT
YEDLEQAFVSDEVSEDALKSCLIDEVNALLEPVRQHFASNEEAHELLEAVKSYRKDGATL
PLAETALPAAPAKPHACMWMPALLKVPLDVAEGMIKVTKDFIAAHPEGTVTLVLPDWSAV
ASDEITGVEKDISAALQVNCALLKAYGLPSSVKIVTENEVILGNCDDFWVSVIGIARKNL
LSHVEELYGGEVRNAGQVIAALMRVATALMLSVSHVISTSLDGHINAFAREYTKERIDCV
QTLEGRIPALHRPGAAPAVLGADDVLYLDDNDMDIRRKIKKAYSAPNEEANPVISVAQHL
LAQHGALSIERGEANGGNVSYNTPEALVADCGSGALHPADLKAAVLQLLLDRSAQARALL
NGELKKNMTVLRNAEKKMAKKR
*length of your protein in Amino Acids: 682aa
Molecular Weight: 74.949 kDa
Molar Extinction coefficient of your protein at 280 nm wavelength: 71945
TMpred graph Image
*CDS Gene Sequence (paste as text only): ATGAACACGGACGAGCGCTACAAGCTCCTCCGCAGCGTCGGAGAGGAGTGCATTCAGGAA
AGTGAGCTGCGCAACCTTATCGAAAAGAAGCCGCTCATTCGCTGCTACGATGGCTTCGAG
CCCTCCGGCCGCATGCACATTGCTCAGGGTATCTTCAAGGCAGTCAATGTCAACAAGTGC
ACCGCCGCCGGGTGCGAGTTTGTCTTCTGGGTGGCGGACTGGTTTGCGCTCATGAACGAC
AAGGTCGGCGGCGAACTGGAGAAGATCCGCATTGTTGGCAGGTACCTTATTGAGGTGTGG
AAGGCGGCGGGCATGGACATGGACAAGGTTTTGTTTTTATGGAGCAGCGAGGAGATCACG
AGCCACGCCGACACGTACTGGCGCATGGTGCTCGACATTGGCCGCCAGAATACGATCGCC
CGCATCAAGAAGTGCTGCACCATCATGGGCAAGACGGAAGGCACGCTGACGGCGGCACAG
GTGCTGTACCCGCTGATGCAGTGCTGCGACATCTTCTTTCTCAAGGCCGACATCTGCCAG
CTGGGCCTGGACCAGCGCAAGGTGAACATGCTCGCACGCGAGTACTGCGATCTGATCGGC
CGCAAGTTGAAGCCGGTCATTCTGTCACACCACATGCTGGCCGGTCTGAAGCAGGGGCAG
GCGAAGATGAGCAAGAGTGACCCGGACAGTGCCATCTTCATGGAAGACACCGAGGAGGAT
GTGGCACGTAAGATCCGTCAGGCCTACTGTCCGCGCGTCAAGCAGTCCGCCAGCGCCATC
ACAGATGACGGGGCGCCGGTCGCCACGGACGACCGCAACCCGGTGCTGGACTACTTCCAG
TGTGTCGTCTACGCCCGACCCGGCGCTGCGGCCACCATTGACGGCACCACCTACGCGACC
TACGAGGACCTCGAGCAGGCTTTTGTGAGCGACGAGGTCAGCGAGGATGCCCTCAAGTCG
TGCCTGATTGATGAGGTGAACGCCCTGCTGGAGCCCGTGCGCCAACATTTTGCGTCAAAC
GAGGAGGCTCACGAGCTGCTGGAGGCGGTGAAGTCGTACCGTAAAGATGGCGCGACCCTT
CCACTGGCCGAGACAGCGCTCCCTGCCGCCCCCGCAAAGCCGCACGCGTGCATGTGGATG
CCGGCGCTGCTGAAGGTGCCGCTGGATGTCGCGGAAGGGATGATCAAAGTCACCAAGGAC
TTCATCGCCGCCCACCCGGAGGGCACGGTGACACTGGTGCTGCCTGACTGGTCGGCGGTG
GCCAGCGACGAAATCACCGGGGTGGAGAAAGACATCTCGGCCGCCCTCCAGGTGAACTGT
GCGCTTCTCAAGGCGTATGGACTGCCGAGCTCGGTGAAGATCGTGACGGAGAACGAGGTC
ATTCTCGGCAACTGCGACGACTTCTGGGTGAGTGTAATCGGCATTGCACGCAAGAACCTG
CTGAGCCACGTCGAGGAGCTGTACGGGGGAGAGGTACGCAACGCTGGTCAGGTCATTGCT
GCCCTCATGCGCGTTGCCACGGCTCTGATGCTATCCGTCTCGCACGTCATCTCAACCTCG
CTGGATGGACACATCAACGCCTTTGCCCGCGAGTACACGAAGGAGCGGATTGACTGCGTG
CAGACGCTGGAGGGACGCATCCCGGCGCTGCACCGCCCTGGCGCCGCCCCAGCGGTGCTC
GGTGCCGACGACGTCCTATACCTGGACGACAATGACATGGACATCCGCCGTAAAATTAAG
AAGGCGTACTCGGCGCCGAACGAGGAGGCCAATCCGGTCATATCGGTTGCCCAGCACCTT
CTCGCACAGCATGGAGCCCTCAGCATCGAGCGCGGGGAGGCCAACGGTGGCAACGTAAGC
TACAACACGCCAGAGGCCCTCGTGGCAGACTGCGGCAGCGGCGCTCTCCACCCTGCCGAC
CTCAAGGCGGCGGTGCTACAGCTGCTGCTGGACCGGTCGGCGCAGGCGCGCGCGCTGCTC
AACGGCGAGCTCAAGAAGAATATGACCGTTCTTCGCAACGCGGAGAAGAAGATGGCAAAG
AAGAGGTAA
*GC% Content for gene:
*CDS Gene Sequence (codon optimized) - copy from output of Primer Design Protocol (paste as text only):
*GC% Content for gene (codon optimized):
Do Not Need this info for Spring (but still copy these lines to your Target page for now)
Primer design results for pNIC-Bsa4 cloning (list seqeunces of all of your ~40 nt long primers):
(link to DNA Works output text file - that should be saved in your Google Docs folder after you did the primer design protocol)
-- Ask a mentor, Dr. B, or a fellow researcher -how to link a GDocs file if you are not sure how to.
Primer design results for 'tail' primers (this is just 2 sequences):